Differential regulation by phosphatidylinositol 4,5-bisphosphate of pituitary plasma-membrane and cytosolic phosphoinositide kinases.
Regulation of phosphatidylinositol kinase (EC 220.127.116.11) and phosphatidylinositol 4-phosphate (PtdIns4P) kinase (EC 188.8.131.52) was investigated in highly enriched plasma-membrane and cytosolic fractions derived from cloned rat pituitary (GH3) cells. In plasma membranes, phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] added exogenously enhanced incorporation of [32P]phosphate from [gamma-32P]MgATP2- into PtdIns(4,5)P2 and PtdIns4P to 150% of control; half-maximal effect occurred with 0.03 mM exogenous PtdIns(4,5)P2. Exogenous PtdIns4P and phosphatidylinositol (PtdIns) had no effect. When plasma membranes prepared from cells prelabelled to isotopic steady state with [3H]inositol were used, there was a MgATP2- dependent increase in the content of [3H]PtdIns(4,5)P2 and [3H]PtdIns4P that was enhanced specifically by exogenous PtdIns(4,5)P2 also. Degradation of 32P- and 3H-labelled PtdIns(4,5)P2 and PtdIns4P within the plasma-membrane fraction was not affected by exogenous PtdIns(4,5)P2. Phosphoinositide kinase activities in the cytosolic fraction were assayed by using exogenous substrates. Phosphoinositide kinase activities in cytosol were inhibited by exogenously added PtdIns(4,5)P2. These findings demonstrate that exogenously added PtdIns(4,5)P2 enhances phosphoinositide kinase activities (and formation of polyphosphoinositides) in plasma membranes, but decreases these kinase activities in cytosol derived from GH3 cells. These data suggest that flux of PtdIns to PtdIns4P to PtdIns(4,5)P2 in the plasma membrane cannot be increased simply by release of membrane-associated phosphoinositide kinases from product inhibition as PtdIns(4,5)P2 is hydrolysed.


Regional interventions to eliminate healthcare-associated infections.
The development of the Health and Human Services (HHS) Action Plan to eliminate healthcare-associated infections (HAIs) and the coordinated efforts of the federal and private sectors to address this patient safety problem present an unprecedented effort to "bend the cost curve" of delivering care through improving quality. Key to this strategy was a multipronged approach at the federal, state, and regional levels. To examine the impact of HHS's Regional Office projects in support of the implementation of the National Action Plan to Eliminate HAIs and to clarify the role played by HHS regions in the multipronged federal effort to combat HAIs. Qualitative; 13 individual semistructured interviews with representatives from 9 regions. Eight interviews were conducted initially and follow-up interviews were conducted 1-year later with original participants. Evaluated results against the modified Context-Input-Process-Product system functions and properties used to evaluate the HAI National Action Plan, including: HAI data and monitoring, knowledge development, infrastructure development, HAI prevention and practice adoption, prioritization, coordination and alignment, accountability and incentives, stakeholder engagement, and resources. Results from the interviews were systematically coded against a framework that documents specific system and properties that characterize the roadmap laid out by the Action Plan. This review assesses progress toward Action Plan goals achieved by these OASH-funded regional projects and finds that these regional activities furthered the National Action Plan goals by addressing the key regional needs. Key to the success of the National Action Plan was the multilevel approach to implementation of initiatives at the federal, regional, and state levels.


Oxidative stress and metabolic pathologies: from an adipocentric point of view.
Oxidative stress plays a pathological role in the development of various diseases including diabetes, atherosclerosis, or cancer. Systemic oxidative stress results from an imbalance between oxidants derivatives production and antioxidants defenses. Reactive oxygen species (ROS) are generally considered to be detrimental for health. However, evidences have been provided that they can act as second messengers in adaptative responses to stress. Obesity represents a major risk factor for deleterious associated pathologies such as type 2 diabetes, liver, and coronary heart diseases. Many evidences regarding obesity-induced oxidative stress accumulated over the past few years based on established correlations of biomarkers or end-products of free-radical-mediated oxidative stress with body mass index. The hypothesis that oxidative stress plays a significant role in the development of metabolic disorders, especially insulin-resistance state, is supported by several studies where treatments reducing ROS production reverse metabolic alterations, notably through improvement of insulin sensitivity, hyperlipidemia, or hepatic steatosis. In this review, we will develop the mechanistic links between oxidative stress generated by adipose tissue in the context of obesity and its impact on metabolic complications development. We will also attempt to discuss potential therapeutic approaches targeting obesity-associated oxidative stress in order to prevent associated-metabolic complications.


Parallel determination of multiple protein metabolite interactions using cell extract, protein microarrays and mass spectrometric detection.
Analysis of interactive networks between proteins and other molecular constituents is of paramount importance to delineate complex cellular processes. In order to facilitate this process, new technologies that allow rapid, high-throughput parallel screening, as well as identification of constituents, are necessary. A particularly powerful combination in this regard could be the use of multiprotein microarrays coupled with mass spectrometry (MS). In the initial step of the method development we applied MS to single-protein microarrays. We demonstrated that even a simplified version of the method allows rapid parallel label-free assay of specific protein interactions with multiple metabolites derived from complex artificial and natural mixtures. The microarrays fabricated by the electrospray deposition technique and cross-linked in glutaraldehyde vapor were brought into contact with droplets of solution containing either a natural extract of baker's yeast cells or an artificial cocktail of metabolites. After washing, the microarrays were placed into 75% methanol to denature proteins and release specifically bound metabolites. The eluates were then analyzed by electrospray ionization mass spectrometry (ESI-MS) to simultaneously detect all the metabolites bound. Such a procedure applied to ten different proteins demonstrated that 50-400 ng of cross-linked protein is enough to obtain ion intensities from metabolites that are well distinguishable above noise. The compatibility of microplates and different microarray designs with MS detection is discussed.


Estimation of plasma uric acid in pregnancy induced hypertension (PIH). Is the test still relevant?
Plasma uric acid and creatinine were estimated at each visit of 59 consenting women attending ante-natal clinic. Results in the third trimester of pregnancy confirmed that the plasma uric acid was able to differentiate between normal pregnancy and those with PIH at P < 0.002. The results of those with ordinary hypertension and the women who developed pre-eclampsia were similar. Plasma creatinine was found to be able to differentiate between women with ordinary hypertension (97.45 +/- 25.03 umol/L) and those with pre-eclampsia (288.75 +/- 191.97 umol/L) at P < 0.01. For patients with pre-eclampsia who developed convulsions, there invariably was a further rise in the plasma uric acid levels. Monitoring of plasma creatinine level among patients with PH will help to predict those at risk of developing pre-eclampsia. In the same way, monitoring of plasma uric acid level in those with pre-eclampsia will help to predict those that will develop eclampsia. The tests are inexpensive but sensitive, which is in contrast to many more expensive electronic alternatives.


Ca2+-sensitive inactivation and facilitation of L-type Ca2+ channels both depend on specific amino acid residues in a consensus calmodulin-binding motif in the(alpha)1C subunit.
L-type Ca(2+) channels are unusual in displaying two opposing forms of autoregulatory feedback, Ca(2+)-dependent inactivation and facilitation. Previous studies suggest that both involve direct interactions between calmodulin (CaM) and a consensus CaM-binding sequence (IQ motif) in the C terminus of the channel's alpha(1C) subunit. Here we report the functional effects of an extensive series of modifications of the IQ motif aimed at dissecting the structural determinants of the different forms of modulation. Although the combined substitution by alanine at five key positions (Ile(1624), Gln(1625), Phe(1628), Arg(1629), and Lys(1630)) abolished all Ca(2+) dependence, corresponding single alanine replacements behaved similarly to the wild-type channel (77wt) in four of five cases. The mutant I1624A stood out in displaying little or no Ca(2+)-dependent inactivation, but clear Ca(2+)- and frequency-dependent facilitation. An even more pronounced tilt in favor of facilitation was seen with the double mutant I1624A/Q1625A: overt facilitation was observed even during a single depolarizing pulse, as confirmed by two-pulse experiments. Replacement of Ile(1624) by 13 other amino acids produced graded and distinct patterns of change in the two forms of modulation. The extent of Ca(2+)-dependent facilitation was monotonically correlated with the affinity of CaM for the mutant IQ motif, determined in peptide binding experiments in vitro. Ca(2+)-dependent inactivation also depended on strong CaM binding to the IQ motif, but showed an additional requirement for a bulky, hydrophobic side chain at position 1624. Abolition of Ca(2+)-dependent modulation by IQ motif modifications mimicked and occluded the effects of overexpressing a dominant-negative CaM mutant.


Shadows of thought: shifting lateralization of human brain electrical patterns during brief visuomotor task.
Dynamic spatial patterns of correlation of electrical potentials recorded from the human brain were shown in diagrams generated by mathematical pattern recognition. The patterns for "move" and "no-move" variants of a brief visuospatial task were compared. In the interval spanning the P300 peak of the evoked potential, higher correlations of the right parietal electrode with occipital and central electrodes distinguished the no-move task from the move task. In the next interval, spanning the readiness potential in the move task, higher correlations of the left central electrode with occipital and frontal electrodes characterized the move task. These results conform to neuropsychological expectations of localized processing and their temporal sequence. The rapid change in the side and site of localized processes may account for conflicting reports of lateralization in studies which lacked adequate spatial and temporal resolution.


[Prophylaxis of postoperative deep vein thromboses by means of weak doses of subcutaneous heparin (author's transl)].
The authors intend to control with the marked fibrinogen test, the real frequency of postoperative deep vein thromboses (DVT) and the eventual prophylactic effect of subcutaneous heparin in weak doses. Heparin was administered 2 hours before the operation and every 8 hours during 6 days thereafter, each time at the dose of 5,000 U subcutaneously. Ninety-nine patients undergoing major surgery were randomized into two groups of 55 control operated patients and 44 operated patients treated with heparin. The radioactive fibrinogen test was used in 27 patients of the control group and in 23 of the treated group. In the control group of 55, 7 DVT were clinically recognized : the isotopic test was positive in 17 patients. In the treated group of 44 no DVT was clinically diagnosed : 6 DVT were recognized by the marked fibrinogen test. There were no noticeable hemorrhagic complications in the treated group. Low dose of subcutaneous heparin is an efficient and safe prophylactic method in postoperative thromboembolic accidents.


The relationship between duty cycle, tachycardia threshold and autostimulation delivery in cardio-responsive vagus nerve stimulation.
Many patients with epilepsy are unable to completely control seizures with medication alone. Vagus nerve stimulation is an effective nonpharmacologic option for treatment-resistant epilepsy. AutoStim technology is a relatively new VNS feature that provides extra stimulations in response to tachycardia, which often precedes seizures. This feature adds a currently unknown number of extra stimulations per day. This study aims to evaluate that actual stimulations' patients receive per day as a function of varying VNS settings. This project was a retrospective chart review. All patients with implanted VNS devices with AutoStim technology managed at Northwestern's epilepsy clinic were included in the study. Data were collected on duty cycle, tachycardia threshold, expected number of stimulations per day, normal mode stimulations per day, and AutoStims per day. Associations between all variables were evaluated. Sixty-seven patients with VNS stimulators met inclusion criteria leading to 305 unique VNS interrogation results. This study demonstrated the smallest number of extra stimulations between tachycardia thresholds of 40 and 60 with absolute minimums at 35 and 45. Our results also demonstrate a trend of decreasing extra stimulations per day with increased duty cycle at each tachycardia threshold evaluated. These data demonstrate the effect of duty cycle and tachycardia threshold on extra stimulations per day and demonstrated the optimal settings for reducing extra stimulations. This research provides reassurance and information to patients and providers about the actual number of stimulations patients receive with AutoStim technology.


Bioluminescence-based detection of microRNA, miR21 in breast cancer cells.
A hybridization assay for the detection of microRNA, miR21 in cancer cells using the bioluminescent enzyme Renilla luciferase (Rluc) as a label, has been developed. MicroRNAs are small RNAs found in plants, animals, and humans that perform key functions in gene silencing and affect early-stage cell development, cell differentiation, and cell death. miRNAs are considered useful early diagnostic and prognostic markers of cancer, candidates for therapeutic intervention, and targets for basic biomedical research. However, methods for highly sensitive and rapid detection of miRNA directly from samples such as cells that can serve as a suitable diagnostics platform are lacking. In that regard, the utilization of the bioluminescent label, Rluc, that offers the advantage of high signal-to-noise ratio, allows for the development of highly sensitive assays for the determination of miRNA in a variety of matrixes. In this paper, we have described the development of a competitive oligonucleotide hybridization assay for the detection of miR21 using the free miR21 and Rluc-labeled miR21 that competes to bind to an immobilized miR21 complementary probe. The miR21 microRNA chosen for this study is of biomedical significance because its levels are elevated in a variety of cancers. Using the optimized assay, a detection limit of 1 fmol was obtained. The assay was employed for the detection of miR21 in human breast adenocarcinoma MCF-7 cells and nontumorigenic epithelial MCF-10A cells. The comparison of miR21 expression level in two cell lines demonstrated higher expression of miR21 in breast cancer cell line MCF-7 compared to the nontumorigenic MCF-10A cells. Further, using the assay developed, the miR21 quantification could be performed directly in cell extracts. The hybridization assay was developed in a microplate format with a total assay time of 1.5 h and without the need for sample PCR amplification. The need for early molecular markers and their detection methods in cancer diagnosis is tremendous. The characteristics of the assay developed in this work show its suitability for early cancer diagnosis based on miRNA as a biomarker.


Preterm meconium staining of the amniotic fluid: associated findings and risk of adverse clinical outcome.
The incidence of preterm meconium staining of the amniotic fluid (MSAF) is uncertain. It may be an indicator of possible listeriosis. It is unclear how great this risk is or whether preterm MSAF is a risk factor for adverse neonatal outcome. To investigate the incidence of preterm MSAF, the incidence of associated maternal and neonatal infection, and the outcomes of the infants at discharge. Retrospective case-control study. Infants < 33 weeks gestation with preterm MSAF born in the Simpson Memorial Maternity Pavilion, Edinburgh between 1 January 1994 and 2 January 2001 were matched with the next infant of the same sex and gestation with clear liquor. Maternal and infant characteristics, culture results, placental histology, and clinical outcomes were compared. Preterm MSAF was observed in 45/1054 (4.3%) infants below 33 weeks gestation. No maternal or infant listeriosis was identified in cases or controls. There was no significant difference in birth weight, Apgar score, or first pH between cases and controls. Preterm MSAF was associated with prolonged rupture of the membranes (odds ratio (OR) 3.34, 95% confidence interval (CI) 1.07 to 10.49), but not maternal hypertension, sepsis, or chorioamnionitis. Severe (grade 3/4) intraventricular haemorrhage was significantly more common in infants with preterm MSAF (OR 2.03, 95% CI 1.62 to 2.53). There was no significant difference in mortality. Early onset sepsis was observed in two cases and three controls. Preterm meconium staining of the amniotic fluid may be associated with increased risk of intraventricular haemorrhage. It does not appear to be a useful indicator of listeriosis.


[Rapid screening of 44 pesticide residues in ginger and scallion by ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry].
An analytical method was established for the simultaneous determination of 44 pesticide residues in ginger and scallion by ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC-Q-TOF/MS). The samples were extracted with acetonitrile containing 0.1% (v/v) acetic acid aqueous solution, and cleaned-up by primary secondary amine (PSA) and octadecyl bonded silica (C18) adsorbents. The compounds were separated on a Poroshell 120 SB-C18 column (100 mm×3.0 mm, 2.7 μm) with 0.1% (v/v) formic acid aqueous solution containing 5 mmol/L ammonium acetate-acetonitrile as mobile phases under gradient elution. The eluent was determined by UPLC-Q-TOF/MS with electrospray ionization in positive mode. The quantification analysis was performed with the external standard method. In all ions MS/MS mode, the compounds were qualitatively screened and confirmed by one data acquisition. The correlation coefficients (r) were greater than 0.995 in the linear ranges of the 44 pesticides. The limits of quantification (LOQs, S/N=10) of the 44 pesticides were 2.5-5.0 μg/kg. At the three spiked levels, the recoveries were between 73.4% and 113.7% with the relative standard deviations (RSDs) ranging from 0.7% to 12.1% (n=6). The method effectively improves the determination efficiency of pesticide residues screening by high-resolution mass spectrometry.


Serological markers and risk factors associated with Hepatitis B virus infection among Federal Capital Territory prison inmates, Nigeria: Should we be concerned?
Hepatitis B virus (HBV) infection is hyper-endemic in Nigeria. Prisons are high-risk environments for the spread of infectious diseases. Worldwide, seroprevalence of HBV infection is substantially higher among individuals in correctional facilities when compared to general population. We determined the seroprevalence and risk factors associated with HBV infection among Kuje prison inmates, Nigeria. We conducted a prison facility based cross-sectional study. Interviewer administered questionnaires were used to obtain information on participants socio-demographic characteristics, HBV risk factors, previous HBV test and vaccination history. Blood samples collected from participants were analysed for HBsAg, HBsAb, HBcAb, HBeAg and HBeAb markers using rapid lateral chromatographic immunoassay kit. Univariate, bivariate, and multivariate analysis were performed. A total of 271 inmates (63 convicts and 208 awaiting trial inmates) were recruited into the study as participants. The mean age of the participants was 32.7 SD±9 years. HBV seroprevalence (HBsAg) of 13.7% (95% CI: 9.8-18.3) was found. 55.4% (95% CI: 49.2-61.4) of inmates were susceptible to HBV infection, 20.7% (95%CI; 16.0-26.0) had past HBV infection while 10.3% (95% CI: 7.0-14.6) had acquired natural or artificial HBV immunity. Factors found to be associated with current HBV infection (HBsAg) include age-group ≤25years (aOR = 8.0,95% CI: 2.9-22.3), being ever married (aOR = 4.2, 95% CI: 1.7-10.4) and history of alcohol consumption (aOR = 3.4, 95% CI: 1.3-8.4). This study reveals a high seroprevalence of HBV infection among Kuje Prison inmates, hence the need to introduce prison-focused health intervention initiatives such as HBV screening, vaccination and care to reduce the transmission of HBV infection among inmates and ultimately the general population.


[New method for measuring the distances between the nitroxide spin label and paramagnetic metal ions in macromolecules].
A new method of estimation of the distance RLM between the nitroxide spin label (NSL) and the paramagnetic metal ions (PMI), such as Co2+, Ni2+, Cu2+, Mn2+, VO2+, Cr3+, Fe3+ is suggested. The influence of the longitudinal relaxation time T1 of the PMI on the line shape of the NSL at 77 degrees K has been studied. It was found that the efficiency of the dipole-dipole interaction between NSL and PMI depends strongly on the T1 value of the PMI. Measurements of the RLM for 4 spin-labelled proteins (haemoglobin, nitrogenase, cytochrome P450 and Ca2+-dependent ATPase) by three various methods have proved the correctness of the new method and also its simplicity.


Rationale for using serum prostate-specific antigen (PSA) level and PSA density (PSAD) to detect prostatic malignancy in a country with low prostate cancer incidence.
Prostate-specific antigen (PSA) and PSA density (PSAD) values in Indonesia had been found uniquely much higher than the normal accepted values in western countries. However, PSA more than 10 ng/ml and PSAD above 0.15 in intermediate PSA of 4-10 ng/ml are still indicative of a prostate biopsy. This condition had led to unnecessary biopsies in view of the low incidence of prostatic carcinoma in our country. Our objective is to find alternative serum PSA levels and PSAD cutoff points that enhance the specificity and sensitivity of prostate cancer detection. This retrospective cross-sectional study included 805 consecutive patients from 40 to 95 years old in Sumber Waras Hospital (SWH) and Cipto Mangunkusumo Hospital (CMH) from 1994 to 1997. All patients underwent digital rectal examination (DRE) and transrectal ultrasonography (TRUS) to measure prostate volume. After the serum PSA level was determined and PSAD was calculated, prostate biopsies were performed if the PSA level was greater than 10 ng/ml or PSAD more than 0.15 at intermediate PSA levels of 4-10 ng/ml. The ability of PSA, intermediate PSA level, and PSAD to improve the accuracy of prostate cancer detection was evaluated using univariate analysis. Among 805 patients, 240 patients had PSA level < 4 ng/ml, 230 patients had PSA level 4-10 ng/ml, and 335 had PSA level > 10 ng/ml. Of the 230 patients with intermediate PSA level, 108 had PSAD > 0.15. Thirty-five patients had histologically confirmed prostatic carcinoma, i.e., 3 of 108 patients with PSA 4-10 ng/ml and PSAD > 0.15, and 32 of 335 patients with PSA > 10 ng/ml. There were 105 and 303 unnecessary biopsies in those groups. With a PSA cutoff level of > or = 8 ng/ml, we found 100% sensitivity to prostate cancer. PSAD > or = 0.20 within a PSA level of 8-30 ng/ml gave 100% sensitivity to prostate cancer. Using these new cutoffs there would be 148 biopsies (33.4%) saved. We concluded from this study that the commonly accepted values of serum PSA level and PSAD resulted in many unnecessary biopsies in our patients. Instead, the most sensitive cutoff points to perform prostate biopsy are serum PSA level greater than 8.0 ng/ml, and PSAD of more than 0.20 at an intermediate serum PSA level of 8-30 ng/ml.


Reality of Family Caregiving for Hispanics With Alzheimer's Disease and Related Dementias: A Qualitative Analysis.
Background: Underserved Hispanic patients often experience unmet palliative care (PC) needs, particularly those with noncancer diagnoses such as Alzheimer's disease and related dementias. Most caregivers for Hispanic patients are family relatives who are less likely to use health care and community resources and experience high caregiver burden. We adapted a culturally tailored patient navigator (PN) intervention to provide support and improve PC outcomes for Hispanics with Alzheimer's disease and related dementias and their family caregivers (FCGs). Objectives: To explore Hispanic FCGs' experiences and perceptions of caregiving for a loved one, and how our PN intervention impacted their needs. Design: Qualitative descriptive. Setting/Subjects: FCG participants (n = 10) from our randomized control trial's intervention group were recruited from academic and safety net hospitals and community-based clinics across urban and rural Colorado in the United States. Measurements: Data obtained from individual, semistructured, 30-minute telephone interviews were recorded, transcribed, translated, and analyzed using NVivo and qualitative thematic analyses. Results: Four major themes emerged: Methods of Support, Cultural Expectations and Varying Family Contributions, Lack of Self-Care, and Awareness. Subthemes highlighted differing definitions of "contributing," role resentment, and interpersonal issues. Varying familial expectations underscore FCG strain when the burden of caregiving is not shared. Participants used various coping strategies as necessary support and gained awareness through education, guidance, and referrals to resources. Conclusions: PNs helped FCGs and patients beyond the intervention's scope. Providing support and awareness to FCGs, and incorporating cultural beliefs, may improve PC access to disparate populations and guide future interventions. Clinical Trial Registration Number NCT03181750.


Multimorbidity, disadvantage, and patient engagement within a specialist homeless health service in the UK: an in-depth study of general practice data.
There is a paucity of current health data regarding users of a specialist homeless health service in the UK. To describe the health of users of a specialist homeless health service by assessing levels of multimorbidity, social exclusion - by measuring severe and multiple disadvantage (SMD) - and patient engagement with health care. Analysis of patient-level data from computerised records of patients registered with a specialist homeless health service in Glasgow, Scotland. Data for 133 patients were extracted using a data extraction form. Multimorbidity and SMD were described using categorisation adapted from previous literature in this field. Stepwise regression analysis was carried out to assess the relationship between domains of SMD experienced and the number of long-term conditions (LTCs) a patient had. The average age of patients in the cohort was 42.8 years, however levels of multimorbidity were comparable to those aged ≥85 years in the general population. The average number of LTCs was 2.8 per patient, with 60.9% of patients having both mental and physical comorbidity. SMD was categorised into three domains: homelessness; substance misuse; and previous imprisonment. More than 90.0% of patients experienced ≥2 domains of SMD, and SMD experiences were associated with multimorbidity: as domains of SMD experiences increased, so did the number of LTCs a patient was recorded as having. This cohort of patients has a complex burden of health and social care needs, which may act as barriers in the provision of effective health care.


Age at natural menopause and risk of incident cardiovascular disease: a pooled analysis of individual patient data.
Early menopause is linked to an increased risk of cardiovascular disease mortality; however, the association between early menopause and incidence and timing of cardiovascular disease is unclear. We aimed to assess the associations between age at natural menopause and incidence and timing of cardiovascular disease. We harmonised and pooled individual-level data from 15 observational studies done across five countries and regions (Australia, Scandinavia, the USA, Japan, and the UK) between 1946 and 2013. Women who had reported their menopause status, age at natural menopause (if postmenopausal), and cardiovascular disease status (including coronary heart disease and stroke) were included. We excluded women who had hysterectomy or oophorectomy and women who did not report their age at menopause. The primary endpoint of this study was the occurrence of first non-fatal cardiovascular disease, defined as a composite outcome of incident coronary heart disease (including heart attack and angina) or stroke (including ischaemic stroke or haemorrhagic stroke). We used Cox proportional hazards models to estimate multivariate hazard ratios (HRs) and 95% CIs for the associations between age at menopause and incident cardiovascular disease event. We also adjusted the model to account for smoking status, menopausal hormone therapy status, body-mass index, and education levels. Age at natural menopause was categorised as premenopausal or perimenopausal, younger than 40 years (premature menopause), 40-44 years (early menopause), 45-49 years (relatively early), 50-51 years (reference category), 52-54 years (relatively late), and 55 years or older (late menopause). Overall, 301 438 women were included in our analysis. Of these 301 438 women, 12 962 (4·3%) had a first non-fatal cardiovascular disease event after menopause, of whom 9369 (3·1%) had coronary heart disease and 4338 (1·4%) had strokes. Compared with women who had menopause at age 50-51 years, the risk of cardiovascular disease was higher in women who had premature menopause (age <40 years; HR 1·55, 95% CI 1·38-1·73; p<0·0001), early menopause (age 40-44 years; 1·30, 1·22-1·39; p<0·0001), and relatively early menopause (age 45-49 years; 1·12, 1·07-1·18; p<0·0001), with a significantly reduced risk of cardiovascular disease following menopause after age 51 years (p<0·0001 for trend). The associations persisted in never smokers, and were strongest before age 60 years for women with premature menopause (HR 1·88, 1·62-2·20; p<0·0001) and early menopause (1·40, 1·27-1·54; p<0·0001), but were attenuated at age 60-69 years, with no significant association observed at age 70 years and older. Compared with women who had menopause at age 50-51 years, women with premature and early menopause had a substantially increased risk of a non-fatal cardiovascular disease event before the age of 60 years, but not after age 70 years. Women with earlier menopause need close monitoring in clinical practice, and age at menopause might also be considered as an important factor in risk stratification of cardiovascular disease for women. Australian National Health and Medical Research Council.


Screening for antimicrobial resistance in fecal samples by the replica plating method.
Replica plating can be used for the detection of antibiotic resistance in normal flora. We have evaluated this application of the replica plating method by comparing it with a five-colony method. The replica plating method uses a single plate for each antibiotic, with a concentration just above that for borderline resistance. By the five-colony method, five colonies per sample were picked, chosen to represent all different colony morphologies present, and MICs were determined by a standard agar dilution method. The gram-negative, aerobic floras of 131 fecal samples were screened for resistance to ampicillin, cefuroxime, nalidixic acid, trimethoprim, sulfamethoxazole, and tetracycline by both methods. The rate of resistance detection by the two methods did not differ statistically for any of the antibiotics tested. The breakpoint concentrations used for the replica plates in the study gave results similar to those produced by the agar dilution method and the breakpoint values of the National Committee for Clinical Laboratory Standards and can thus be recommended. As the only currently used resistance detection method, replica plating facilitates an exact determination of the percentage of resistant colonies/total number of colonies (between 1 and 100%) in a sample. This revealed an uneven distribution, with only 23% of the samples having resistance frequencies in the range of 10 to 85%; usually, the resistant flora either was a small minority or was very dominant in samples with resistance. This phenomenon was present for all of the antibiotics.


[Contribution of glucocorticoid hormones to gastroprotection].
The fact that the disturbance of the normal stress reaction by the elimination of the hypothalamic-pituitary-adrenocortical (HPA) axis's functioning leads to negative effects on the body such as the development and aggravation of diseases proves that stress plays a leading role in maintaining the physical health of the body. Here we demonstrate this on the base of the results of our experimental studies related with gastric ulceration. The results show that the reduction in the stress-induced corticosterone release, or its actions, aggravates stress-caused gastric erosion. The data do not support the traditional paradigm about ulcerogenic role of glucocorticoids in stress and suggest that glucocorticoids released during acute activation of the HPA axis are naturally occurring gastroprotective factors.


Low paternal dietary folate alters the mouse sperm epigenome and is associated with negative pregnancy outcomes.
Epidemiological studies suggest that a father's diet can influence offspring health. A proposed mechanism for paternal transmission of environmental information is via the sperm epigenome. The epigenome includes heritable information such as DNA methylation. We hypothesize that the dietary supply of methyl donors will alter epigenetic reprogramming in sperm. Here we feed male mice either a folate-deficient or folate-sufficient diet throughout life. Paternal folate deficiency is associated with increased birth defects in the offspring, which include craniofacial and musculoskeletal malformations. Genome-wide DNA methylation analysis and the subsequent functional analysis identify differential methylation in sperm of genes implicated in development, chronic diseases such as cancer, diabetes, autism and schizophrenia. While >300 genes are differentially expressed in offspring placenta, only two correspond to genes with differential methylation in sperm. This model suggests epigenetic transmission may involve sperm histone H3 methylation or DNA methylation and that adequate paternal dietary folate is essential for offspring health.


The influence of 'time since last blood meal' on the toxicity of essential oils to the poultry red mite (Dermanyssus gallinae).
The poultry red mite, Dermanyssus gallinae (De Geer) is a serious ectoparasitic pest of layer hens that can survive for long periods in the poultry house sub-structure without taking a blood meal from its host. The research undertaken in this study found that 'time since last blood meal' had a notable effect on how toxic a selection of plant essential oils were to D. gallinae under laboratory conditions. In general, the essential oils had a greater toxic effect on D. gallinae if mites had been starved of a blood meal for around 3 weeks, than if they had been more recently fed 3-13 days prior to tests. This result was consistent across the four essential oils used (thyme, palmarosa, caraway and juniper leaf). This suggests that plant essential oils may be of use in management schemes for D. gallinae, particularly if used to sanitise houses between flocks, when mites will have been starved.


Structural dynamics effects on the ultrafast chemical bond cleavage of a photodissociation reaction.
The correlation between chemical structure and dynamics has been explored in a series of molecules with increasing structural complexity in order to investigate its influence on bond cleavage reaction times in a photodissociation event. Femtosecond time-resolved velocity map imaging spectroscopy reveals specificity of the ultrafast carbon-iodine (C-I) bond breakage for a series of linear (unbranched) and branched alkyl iodides, due to the interplay between the pure reaction coordinate and the rest of the degrees of freedom associated with the molecular structure details. Full-dimension time-resolved dynamics calculations support the experimental evidence and provide insight into the structure-dynamics relationship to understand structural control on time-resolved reactivity.


Selective fine needle aspiration of parotid masses. FNA should be performed in all patients older than 60 years.
The exact role of fine needle aspiration in the pre-operative assessment of patients presenting with parotid masses is controversial. Some surgeons propose that fine needle aspiration be performed only selectively in those patients with likely malignant disease, whilst others recommend it for all patients presenting with such a mass. Intuitively, one would expect older patients to be more likely to suffer from primary malignant parotid tumours and secondary deposits of malignant skin tumours. Therefore, we hypothesised that older patients with a parotid mass should undergo fine needle aspiration regardless of their medical history. We retrospectively reviewed 197 consecutive parotidectomies to test this hypothesis. One hundred and twenty-one patients (61.4 per cent) were diagnosed with benign disease, whilst 76 (38.6 per cent) were diagnosed with malignant disease. Eighty-three per cent of patients aged 60 years or younger had benign disease, as opposed to 35.6 per cent of patients aged more than 60 years. Malignant disease occurred more commonly in patients older than 60 years (odds ratio 8.962, 95 per cent confidence interval 4.607-17.434). In patients with a parotid mass, fine needle aspiration should be performed on all those aged 60 years or older.


MicroRNAs as potential biomarkers for the diagnosis of Traumatic Brain Injury: A systematic review and meta-analysis.
Background: Traumatic brain injury (TBI) is a sudden trauma on the head, in which severe TBI (sTBI) is usually associated with death and long-term disability. MicroRNAs (miRNAs) are potential biomarkers of diverse diseases, including TBI. However, few systematic reviews and meta-analyses have been conducted to determine the clinical value of miRNAs expression in TBI patients. Methods: We conducted this systematic review and meta-analysis study according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We searched PubMed, Embase, the Cochrane Library, Web of Science, from inception to August 26, 2020. We included articles written in English that have reported on the diagnostic value of miRNAs expression in TBI patients. We excluded studies that did not provided sufficient information to construct the 2×2 contingency table. Results: Eight studies investigating the diagnostic value of miRNA in TBI were analyzed in this study. The overall sensitivity, specificity and area under the curve (AUC) of miRNAs in diagnosis of TBI were 89% [95% confidence interval (CI): 0.84-0.93], 92% (95% CI 0.82-0.97) and 95% (95% CI 0.93-0.97). We found that panels of multiple miRNAs could improve the diagnostic accuracy of TBI. Samples from blood and brain tissue have significantly enhanced diagnostic accuracy, when compared with saliva. The AUC of miRNAs in severe TBI was 0.97, with 91% sensitivity and 92% specificity. Conclusion: This systematic review and meta-analysis demonstrated that miRNAs could be potential diagnostic markers in TBI patients. MiRNAs detected in blood and brain tissue display high accuracy for TBI diagnosis.


Long-term follow-up after radical cystectomy with emphasis on complications and reoperations: a Swedish population-based survey.
To evaluate outcome after radical cystectomy for primary bladder cancer in a large population-based material. Between 1997 and 2002 all patients treated with radical cystectomy within 3 months after diagnosis of primary bladder cancer without distant metastasis were retrieved through the Swedish Bladder Cancer Registry. A follow-up questionnaire was distributed to all units where the primary registration of patients was performed. Follow-up data on recurrence date were retrieved from the patient charts and causes of death were obtained from the Swedish Cause of Death Registry until 2003. During the study period radical cystectomy was performed in 39 units in Sweden, of which only five units were considered high-volume hospitals performing 10 or more procedures annually. Mean blood loss was 2300 ml (median 2000 ml) and the 90-day mortality rate was 5.7%. Blood loss was higher in high-volume units than in hospitals with lower hospital volumes, but the 90-day mortality rates were similar. During a median follow-up of 3.5 years, 24% of the patients were submitted to a reoperation. Reoperation rates were significantly higher in patients who received a continent urinary diversion (29%) compared with an ileal conduit (22%, p < 0.015). Radical cystectomy was associated with a reoperation rate of 24% in Sweden during the study period. The reoperation rates were higher in patients receiving a continent cutaneous diversion or bladder substitution. Blood loss was higher in high-volume units; otherwise, surgical volume did not affect mortality rates, cancer-specific survival or reoperation rates.


Protein trafficking abnormalities in Drosophila tissues with impaired activity of the ZIP7 zinc transporter Catsup.
Developmental patterning requires the precise interplay of numerous intercellular signaling pathways to ensure that cells are properly specified during tissue formation and organogenesis. The spatiotemporal function of the Notch signaling pathway is strongly influenced by the biosynthesis and intracellular trafficking of signaling components. Receptors and ligands must be trafficked to the cell surface where they interact, and their subsequent endocytic internalization and endosomal trafficking is crucial for both signal propagation and its down-modulation. In a forward genetic screen for mutations that alter intracellular Notch receptor trafficking in Drosophila epithelial tissues, we recovered mutations that disrupt the Catsup gene, which encodes the Drosophila ortholog of the mammalian ZIP7 zinc transporter. Loss of Catsup function causes Notch to accumulate abnormally in the endoplasmic reticulum (ER) and Golgi compartments, resulting in impaired Notch signaling. In addition, Catsup mutant cells exhibit elevated ER stress, suggesting that impaired zinc homeostasis causes increased levels of misfolded proteins within the secretory compartment.


Feeding Unprotected CLA Methyl Esters Compared to Sunflower Seeds Increased Milk CLA Level but Inhibited Milk Fat Synthesis in Cows.
An experiment was conducted to compare the effect of the same amount of 18:2 offered either as 18:2n-6 or as a mixture of unprotected 18:2c9t11 and 18:2t10c12 on feed intake, milk components as well as plasma and milk fatty acid profile. Fifteen cows were blocked by milk yield and milk fat percentage and within block assigned randomly to 1 of 3 treatments (n = 5). Each cow passed a 12-d adjustment period (AP) on a basal diet. After the AP cows received 1 of 3 supplements during an 18-d experimental period (EP). The supplements contained either 1.0 kg ground sunflower seeds (S), 0.5 kg conjugated linoleic acid (CLA)-oil (C) or 0.75 kg of a mixture of ground sunflower seeds and CLA-oil (2:1; SC). All 3 supplements contained the same amount of 18:2 either as CLA (∑18:2c9t11+18:2t10c12, 1:1) or as 18:2c9c12. During the last 2 d of AP and the last 4 d of EP feed intake and milk yield were recorded daily and milk samples were collected at each milking. Blood samples were collected from the jugular vein on d 11 of AP and d 15 and 18 of EP. The 18:2 intake increased in all treatments from AP to EP. Regardless of the amount of supplemented CLA, the milk fat percentage decreased by 2.35 and 2.10%-units in treatment C and SC, respectively, whereas in the treatment S the decrease was with 0.99%-unit less pronounced. Thus, C and SC cows excreted daily a lower amount of milk fat than S cows. The concentration of trans 18:1 in the plasma and the milk increased from AP to EP and increased with increasing dietary CLA supply. While the concentration of 18:2c9t11 and 18:2t10c12 in the plasma and that of 18:2t10c12 in the milk paralleled dietary supply, the level of 18:2c9t11 in the milk was similar in C and CS but still lower in S. Although the dietary concentration of CLA was highest in treatment C, the partial replacement of CLA by sunflower seeds had a similar inhibitory effect on milk fat synthesis. Comparable 18:2c9t11 levels in the milk in both CLA treatments implies that this isomer is subjected to greater biohydrogenation with increasing supply than 18:2t10c12. The fact that unprotected 18:2t10c12 escaped biohydrogenation in sufficient amounts to affect milk fat synthesis reveals opportunities to develop feeding strategies where reduced milk fat production is desirable or required by the metabolic state of the cow.


Developmental effects of dietary phytoestrogens in Sprague-Dawley rats and interactions of genistein and daidzein with rat estrogen receptors alpha and beta in vitro.
Estrogenic isoflavones, such as genistein and daidzein, are present in virtually all natural-ingredient rodent diets that use soy as a source of protein. Since these compounds are endocrine-active, it is important to determine whether the amounts present in rodent diets are sufficient to affect sexual development. The present study consisted of in vitro and in vivo parts. In the in vitro portion, human hepatoma cells were transfected with either rat estrogen receptor (ER) alpha or beta plus an estrogen-responsive luciferase reporter gene. Genistein and daidzein were complete agonists at both ERs, genistein being more potent than daidzein, and both compounds were more potent at ER beta than ER alpha. In combined studies with estradiol, genistein exerted additive effects with estradiol in vitro. In the in vivo portion of the study, groups of six pregnant Sprague-Dawley females were fed one of the following four diets, and the pups were maintained on the same diets until puberty: (1) a natural-ingredient, open-formula rodent diet (NIH-07) containing 16 mg genistein and 14 mg daidzein per 100 g of feed; (2) a soy- and alfalfa-free diet (SAFD) in which casein and corn oil were substituted for soy and alfalfa meal and soy oil, respectively, that contained no detectable isoflavones; (3) SAFD containing 0.02% genistein (GE.02); or (4) SAFD containing 0.1% genistein (GE.1). In the GE.1 group, effects of dietary genistein included a decreased rate of body-weight gain, a markedly increased (2.3-fold) uterine/body weight (U/BW) ratio on postnatal day (pnd) 21, a significant acceleration of puberty among females, and a marginal decrease in the ventral prostate weight on postnatal day (pnd) 56. However, developmental differences among the groups fed SAFD, GE.02, or NIH-07 were small and suggested minimal effects of phytoestrogens at normal dietary levels. In particular, on pnd 21, the U/BW ratio of the GE.02 and NIH-07 groups did not differ significantly from that of the SAFD group. Only one statistically significant difference was detected between groups fed SAFD and NIH-07: the anogenital distance (AGD) of female neonates on pnd 1 whose dams were fed NIH-07 was 12% larger than that of neonates whose dams were fed SAFD. The results suggest that normal amounts of phytoestrogens in natural-ingredient rodent diets may affect one developmental parameter, the female AGD, and that higher doses can affect several other parameters in both males and females. Based on these findings, we do not suggest replacing soy- and alfalfa-based rodent diets with phytoestrogen-free diets in most developmental toxicology studies. However, phytoestrogen-free diets are recommended for endocrine toxicology studies at low doses, to determine whether interactive effects may occur between dietary phytoestrogens and man-made chemicals.


The Biochemical Mechanism of Fork Regression in Prokaryotes and Eukaryotes-A Single Molecule Comparison.
The rescue of stalled DNA replication forks is essential for cell viability. Impeded but still intact forks can be rescued by atypical DNA helicases in a reaction known as fork regression. This reaction has been studied at the single-molecule level using the Escherichia coli DNA helicase RecG and, separately, using the eukaryotic SMARCAL1 enzyme. Both nanomachines possess the necessary activities to regress forks: they simultaneously couple DNA unwinding to duplex rewinding and the displacement of bound proteins. Furthermore, they can regress a fork into a Holliday junction structure, the central intermediate of many fork regression models. However, there are key differences between these two enzymes. RecG is monomeric and unidirectional, catalyzing an efficient and processive fork regression reaction and, in the process, generating a significant amount of force that is used to displace the tightly-bound E. coli SSB protein. In contrast, the inefficient SMARCAL1 is not unidirectional, displays limited processivity, and likely uses fork rewinding to facilitate RPA displacement. Like many other eukaryotic enzymes, SMARCAL1 may require additional factors and/or post-translational modifications to enhance its catalytic activity, whereas RecG can drive fork regression on its own.


Conditional transgene expression mediated by the mouse beta-actin locus.
Transgenic mice are an effective model to study gene function in vivo; however, position effects can complicate tissue-specific transgene analysis. To facilitate precise targeting of a transgenic construct into the mouse genome, we combined the Cre/lox and Flp/FRT recombination systems to allow for rapid transgene replacement and conditional transgene expression from the endogenous beta-actin locus. Flp/FRT recombination was used to rapidly exchange FRT-flanked transgene cassettes by recombinase-mediated cassette exchange in embryonic stem cells, while transgene expression can be activated in mice after Cre-mediated excision of a floxed STOP cassette. To validate our system, we analyzed the expression profile of an EGFP reporter gene after integration into the beta-actin locus and Cre-mediated excision of the floxed STOP cassette. Breeding of EGFP reporter mice with various Cre mouse lines resulted in the expected expression profiles, demonstrating the feasibility of the model to facilitate predictable and strong transgene expression in a spatially and temporally controlled manner.


Regional differences in quantities of histochemically detectable mucosubstances in nasal, paranasal, and nasopharyngeal epithelium of the bonnet monkey.
Inhaled irritants induce secretory cell hyperplasia in nasal epithelium of animals. To characterize this response histochemically it is first important to know the histochemical character and distribution of epithelial mucosubstance in the normal nasal cavity. An automated image analyzing method was used to detect and quantitate acidic, neutral, and sulfated mucosubstances in the epithelium lining the nasal and paranasal airways of eight bonnet monkeys. Tissue sections 2 micron thick from defined regions of these airways were stained with either alcian blue/periodic acid-Schiff to demonstrate acid and neutral mucosubstances or high iron diamine to demonstrate sulfated mucins. Respiratory epithelium covering maxilloturbinates had the largest volume of stainable mucosubstance per unit surface area of basal lamina, whereas the maxillary sinus epithelium had the least. There was a general anteroposterior increase in the quantity of total epithelial mucosubstance along the septal and lateral walls of the nasal cavity, and there was more acidic than neutral mucosubstance in the posterior nasal airway than in the anterior. Epithelial mucosubstance in the maxillary sinus was predominantly neutral. Therefore, we conclude that there are substantial regional quantitative differences in stainable mucosubstances in the primate nasal epithelium which must be considered when examining nasal mucosa for irritant-induced changes in epithelial mucins.


A genetical analysis of covariation between finger ridge counts.
1. The genetical and environmental structure of covariation between finger ridge counts in twin and sibling data has been analysed using the method of Martin and Eaves (1977) adapted from Jöreskog. 2. The model for environmental covariance contains a single factor loading on all ten digits but most environmental variance is specific to each finger. 3. For additive genetic variance there is one common factor loading on all digits. There are also five other independent factors, one for each digit. The thumb factor loads only on the two thumbs but the four fingers load on the finger in question and on the adjacent fingers. 4. A single common factor for non-additive genetic variance produces a considerable improvement in the model. 5. The pattern of genetic effects differs between left and right hands. 6. Although the same model is appropriate for males and females, different parameters estimates are required. 7. The fit of models is sensitive to the scale of measurement.


Combinatorial ECM Arrays Identify Cooperative Roles for Matricellular Proteins in Enhancing the Generation of TH+ Neurons From Human Pluripotent Cells.
The development of efficient cell culture strategies for the generation of dopaminergic neurons is an important goal for transplantation-based approaches to treat Parkinson's disease. To identify extracellular matrix molecules that enhance differentiation and might be used in these cell cultures we have used micro-contact printed arrays on glass slides presenting 190 combinations of 19 extracellular matrix molecules selected on the basis of their expression during embryonic development of the ventral midbrain. Using long-term neuroepithelial stem cells (Lt-NES), this approach identified a number of matricellular proteins that enhanced differentiation, with the combination of Sparc, Sparc-like (Sparc-l1) and Nell2 increasing the number of tyrosine hydroxylase+ neurons derived from Lt-NES cells and, critically for further translation, human pluripotent stem cells.


Collective attacks and unconditional security in continuous variable quantum key distribution.
We present here an information theoretic study of Gaussian collective attacks on the continuous variable key distribution protocols based on Gaussian modulation of coherent states. These attacks, overlooked in previous security studies, give a finite advantage to the eavesdropper in the experimentally relevant lossy channel, but are not powerful enough to reduce the range of the reverse reconciliation protocols. Secret key rates are given for the ideal case where Bob performs optimal collective measurements, as well as for the realistic cases where he performs homodyne or heterodyne measurements. We also apply the generic security proof of Christiandl et al. to obtain unconditionally secure rates for these protocols.


Potential mechanisms linking atherosclerosis and increased cardiovascular risk in COPD: focus on Sirtuins.
The development of atherosclerosis is a multi-step process, at least in part controlled by the vascular endothelium function. Observations in humans and experimental models of atherosclerosis have identified monocyte recruitment as an early event in atherogenesis. Chronic inflammation is associated with ageing and its related diseases (e.g., atherosclerosis and chronic obstructive pulmonary disease). Recently it has been discovered that Sirtuins (NAD+-dependent deacetylases) represent a pivotal regulator of longevity and health. They appear to have a prominent role in vascular biology and regulate aspects of age-dependent atherosclerosis. Many studies demonstrate that SIRT1 exhibits anti-inflammatory properties in vitro (e.g., fatty acid-induced inflammation), in vivo (e.g., atherosclerosis, sustainment of normal immune function in knock-out mice) and in clinical studies (e.g., patients with chronic obstructive pulmonary disease). Because of a significant reduction of SIRT1 in rodent lungs exposed to cigarette smoke and in lungs of patients with chronic obstructive pulmonary disease (COPD), activation of SIRT1 may be a potential target for chronic obstructive pulmonary disease therapy. We review the inflammatory mechanisms involved in COPD-CVD coexistence and the potential role of SIRT1 in the regulation of these systems.


Rapid access to new bioconjugates of betulonic acid via click chemistry.
Plant-derived pentacyclic triterpenoids of lupane and oleanane families provide a versatile structural platform for the discovery of new biologically active compounds. A number of semisynthetic derivatives of these molecules, possess high medical efficiency including antiviral (HIV-1), anticancer and immunomodulating activity. Even small structural changes in these triterpenoid derivatives were reported to lead to significant changes in their activity, making a convincing case for a systematic study of structure-activity relationships in this class of compounds. Our earlier work opened synthetic access to alkynes derived from the betulonic scaffold and enabled the development of a new family of biohybrids using Click Chemistry (CC). The computer-aided prediction of several types of biological activity were performed with program PASS (Prediction Activity Spectra of Substances. Experimental studies based on mouse models verified the SAR predictions obtained by the PASS program. The observed correlation between the anti-inflammatory and antioxidant activity indicates substantial contribution of the latter in the mechanism of anti-inflammatory effect of the triazole derivatives of betulonic acid.


Prevalence and outcomes of diabetes, hypertension and cardiovascular disease in COPD.
Chronic obstructive pulmonary disease (COPD) is associated with important chronic comorbid diseases, including cardiovascular disease, diabetes and hypertension. The present study analysed data from 20,296 subjects aged > or =45 yrs at baseline in the Atherosclerosis Risk in Communities Study (ARIC) and the Cardiovascular Health Study (CHS). The sample was stratified based on baseline lung function data, according to modified Global Initiative for Obstructive Lung Disease (GOLD) criteria. Comorbid disease at baseline and death and hospitalisations over a 5-yr follow-up were then searched for. Lung function impairment was found to be associated with more comorbid disease. In logistic regression models adjusting for age, sex, race, smoking, body mass index and education, subjects with GOLD stage 3 or 4 COPD had a higher prevalence of diabetes (odds ratio (OR) 1.5, 95% confidence interval (CI) 1.1-1.9), hypertension (OR 1.6, 95% CI 1.3-1.9) and cardiovascular disease (OR 2.4, 95% CI 1.9-3.0). Comorbid disease was associated with a higher risk of hospitalisation and mortality that was worse in people with impaired lung function. Lung function impairment is associated with a higher risk of comorbid disease, which contributes to a higher risk of adverse outcomes of mortality and hospitalisations.


Clinical characteristics and outcomes of chédiak-Higashi syndrome: a nationwide survey of Japan.
Chédiak-Higashi syndrome (CHS) is a rare autosomal recessive disorder characterized by immunodeficiency, neurological dysfunction, and oculocutaneous albinism. Recently, several clinical CHS phenotypes have been reported. Here, we report results of a nationwide survey performed to clarify clinical characteristics and outcomes of CHS patients in Japan. Questionnaires were sent to 287 institutions to collect data regarding CHS patients diagnosed between 2000 and 2010, including results of lysosomal trafficking regulator (LYST) gene analysis. Cytotoxicity and degranulation activity of cytotoxic T lymphocytes were analyzed in available patient samples. A total of 15 patients diagnosed with CHS were eligible for enrollment in this study. Of these, 10 (67%) had recurrent bacterial infections, five (33%) developed life-threatening hemophagocytic lymphohistiocytosis (HLH), and one patient had complicated malignant lymphoma. Hematopoietic stem cell transplantation (HSCT) was performed for six patients including three with HLH, and 10 of the enrolled patients have survived at the time of this writing. LYST analysis was performed for 10 patients; seven different mutations were detected in seven patients, whereas no mutation was identified in three patients. Cytotoxicity and degranulation activity were impaired in patients with and without LYST mutation. Results of this survey indicate that one or two patients with CHS were newly diagnosed each year in Japan. The incidence of HLH was not as high as expected. Mutations of genes other than LYST were suspected in some cases. We conclude that determining indication for HSCT for CHS patients should be based on genetic and cytotoxic analysis.


In Vitro Fertilization Informed Consent: Revisited, Empirically.
A thorough informed consent (IC) process is required before in vitro fertilization (IVF) treatments can begin because these treatments are by and large elective and they have expectable and preventable complications, such as ovarian hyper-stimulation syndrome and multi-fetal pregnancies. To empirically examine whether patient knowledge and understanding of potential hazards associated with IVF treatment are better after the IC process compared to before. The authors hypothesized that patients' better understanding of potential complications would be translated and expressed as rational choices of treatment alternatives. Responses of 48 IVF patients after IC process (study group) from two IVF units in northern Israel were compared to those of 46 patients before IVF (control group). Only women undergoing IVF for first time who were older than 18 years of age were eligible for the study. Socio-demographic parameters were found to be quite similar between the study group and the control group. Contrary to our expectations, in the study group 12 women (25.5%) considered delivery of a single baby as their optimal result, compared to 15 (32.6%) in the control group. Furthermore, preferences shifted toward triplets: eight patients (17%) after IC considered this option as their best result, compared to only five patients (11%) before IC. C process goals are not achieved under current practices, at least as far as IVF treatment are concerned. New tools and incentives should be implemented to meet the requirements dictated by the laws regarding patient rights.


Adult small bowel obstruction.
Small bowel obstruction (SBO) is a clinical condition that is often initially diagnosed and managed in the emergency department (ED). The high rates of potential complications that are associated with an SBO make it essential for the emergency physician (EP) to make a timely and accurate diagnosis. The primary objective was to perform a systematic review and meta-analysis of the history, physical examination, and imaging modalities associated with the diagnosis of SBO. The secondary objectives were to identify the prevalence of SBO in prospective ED-based studies of adult abdominal pain and to apply Pauker and Kassirer's threshold approach to clinical decision-making to the diagnosis and management of SBO. MEDLINE, EMBASE, major emergency medicine (EM) textbooks, and the bibliographies of selected articles were scanned for studies that assessed one or more components of the history, physical examination, or diagnostic imaging modalities used for the diagnosis of SBO. The selected articles underwent a quality assessment by two of the authors using the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2) tool. Data used to compile sensitivities and specificities were obtained from these studies and a meta-analysis was performed on those that examined the same historical component, physical examination technique, or diagnostic test. Separate information on the prevalence and management of SBO was used in conjunction with the meta-analysis findings of computed tomography (CT) to determine the test and treatment threshold. The prevalence of SBO in the ED was determined to be approximately 2% of all patients who present with abdominal pain. Having a previous history of abdominal surgery, constipation, abnormal bowel sounds, and/or abdominal distention on examination were the best history and physical examination predictors of SBO. X-ray was determined to be the least useful imaging modality for the diagnosis of SBO, with a pooled positive likelihood ratio (+LR) of 1.64 (95% confidence interval [CI] = 1.07 to 2.52). On the other hand, CT and magnetic resonance imaging (MRI) were both quite accurate in diagnosing SBO with +LRs of 3.6 (5- to 10-mm slices, 95% CI = 2.3 to 5.4) and 6.77 (95% CI = 2.13 to 21.55), respectively. Although limited to only a select number of studies, the use of ultrasound (US) was determined to be superior to all other imaging modalities, with a +LR of 14.1 (95% CI = 3.57 to 55.66) and a negative likelihood ratio (-LR) of 0.13 (95% CI = 0.08 to 0.20) for formal scans and a +LR of 9.55 (95% CI = 2.16 to 42.21) and a -LR of 0.04 (95% CI = 0.01 to 0.13) for beside scans. Using the CT results of the meta-analysis for the 5- to 10-mm slice subgroup as well as information on intravenous (IV) contrast reactions and nasogastric (NG) intubation management, the pretest probability threshold for further testing was determined to be 1.5%, and the pretest probability threshold for beginning treatment was determined to be 20.7%. The potentially useful aspects of the history and physical examination were limited to a history of abdominal surgery, constipation, and the clinical examination findings of abnormal bowel sounds and abdominal distention. CT, MRI, and US are all adequate imaging modalities to make the diagnosis of SBO. Bedside US, which can be performed by EPs, had very good diagnostic accuracy and has the potential to play a larger role in the ED diagnosis of SBO. More ED-focused research into this area will be necessary to bring about this change.


An In Vitro Study of Three Types of Pit and Fissure Sealants for Viscosity, Resin Tag, and Microleakage: A Scanning Electron Microscope Study.
One of the important techniques for prevention of dental caries is the application of pit and fissure sealants. The sealant success depends on not only adhering firmly to the enamel surface, but also isolation of pits and fissures from the rest of the oral environment.The study was conducted to evaluate and compare three pit and fissure sealant materials for viscosity, microleakage, and resin tag length. Sixty premolars were selected for the study. The teeth were randomly divided into three groups. Group I: type VII GIC, group II: Delton FS, group III: Delton. Teeth were prepared by pumice prophylaxis and etched with 37% phosphoric acid. The respective pit and fissure sealants were applied. Teeth were stored in 1% methylene blue dye and sectioned mesiodistally into two halves. The microleakage was assessed using stereomicroscope and resin tag length using scanning electron microscope (SEM). Brookfield's viscometer was used to measure the viscosity. Viscosity was lowest for Delton and highest for type VII GIC. Microleakage was seen least with the Delton (0.0%) sealant, followed by Delton FS (20.0%), and highest in Type VII GIC (85.0%) in the scoring criteria of 3 and 4 and the mean length of the resin tags was least for Type VII GIC was 3.79 ± 2.58 µm, followed by Delton FS was 5.60 ± 2.12 µm, and highest for Delton was 7.27 ± 3.43 µm. The relation between viscosity, resin tag length, and microleakage was negative. As the viscosity gets lower, the more long the resin tags formed and the microleakage decreased. Delton pit and fissure sealant had lowest viscosity and microleakage scores while longest resin tag lengths. Delton with the lowest viscosity and microleakage appears to be a more suitable pit and fissure sealant compatible with residual moisture and ideal for use in children, where isolation is a problem. Singh R, Lakhanam M. An In Vitro Study of Three Types of Pit and Fissure Sealants for Viscosity, Resin Tag, and Microleakage: A Scanning Electron Microscope Study. Int J Clin Pediatr Dent 2022;15(3):304-310.


Bradykinin does not mediate remote ischaemic preconditioning or ischaemia-reperfusion injury in vivo in man.
To examine whether endogenous bradykinin mediates the endothelium-dependent vasomotor dysfunction induced by ischaemia-reperfusion injury, or the protection afforded by remote ischaemic preconditioning in vivo in man. Randomised double-blind, cross-over study. Royal Infirmary of Edinburgh, Wellcome Trust Clinical Research Facility. Twenty healthy male volunteers. Subjects were randomised to intravenous infusion of the bradykinin B(2) receptor antagonist, HOE-140 (100 μg/kg), or saline placebo in a double-blind, crossover trial. Ischaemia-reperfusion injury was induced in the non-dominant arm by inflating a cuff to 200 mm Hg for 20 min in all subjects. Ischaemia-reperfusion injury was preceded by three cycles of remote ischaemic preconditioning in the dominant arm in 10 subjects. Bilateral forearm blood flow was assessed using venous occlusion plethysmography during intra-arterial infusion of acetylcholine (5-20 μg/min). Acetylcholine caused vasodilatation in all studies (p<0.05) that was attenuated by ischaemia-reperfusion injury, both in the presence (p=0.0002) and absence (p=0.04) of HOE-140. Remote ischaemic preconditioning abolished the impairment of endothelium-dependent vasomotor function induced by ischaemia-reperfusion injury. HOE-140 had no effect on the protection afforded by remote ischaemic preconditioning. These findings do not support a major role for endogenous bradykinin, acting via the B(2) kinin receptor, in the mechanism of ischaemia-reperfusion injury or the protective effects of remote ischaemic preconditioning in man. NCT00965120 and NCT00965393.


A Cascade-Reaction Nanoreactor Composed of a Bifunctional Molecularly Imprinted Polymer that Contains Pt Nanoparticles.
This study was aimed at addressing the present challenge of cascade reactions, namely, how to furnish the catalysts with desired and hierarchical catalytic ability. This issue was addressed by constructing a cascade-reaction nanoreactor made of a bifunctional molecularly imprinted polymer containing acidic catalytic sites and Pt nanoparticles. The acidic catalytic sites within the imprinted polymer allowed one specified reaction, whereas the encapsulated Pt nanoparticles were responsible for another coupled reaction. To that end, the unique imprinted polymer was fabricated by using two well-coupled templates, that is, 4-nitrophenyl acetate and 4-nitrophenol. The catalytic hydrolysis of the former compound at the acidic catalytic sites led to the formation of the latter compound, which was further reduced by the encapsulated Pt nanoparticles to 4-aminophenol. Therefore, this nanoreactor demonstrated a catalytic-cascade ability. This protocol opens up the opportunity to develop functional catalysts for complicated chemical processes.


Rates of major depressive disorder and clinical outcomes following traumatic brain injury.
Uncertainties exist about the rates, predictors, and outcomes of major depressive disorder (MDD) among individuals with traumatic brain injury (TBI). To describe MDD-related rates, predictors, outcomes, and treatment during the first year after TBI. Cohort from June 2001 through March 2005 followed up by structured telephone interviews at months 1 through 6, 8, 10, and 12 (data collection ending February 2006). Harborview Medical Center, a level I trauma center in Seattle, Washington. Five hundred fifty-nine consecutively hospitalized adults with complicated mild to severe TBI. The Patient Health Questionnaire (PHQ) depression and anxiety modules were administered at each assessment and the European Quality of Life measure was given at 12 months. Two hundred ninety-seven of 559 patients (53.1%) met criteria for MDD at least once in the follow-up period. Point prevalences ranged between 31% at 1 month and 21% at 6 months. In a multivariate model, risk of MDD after TBI was associated with MDD at the time of injury (risk ratio [RR], 1.62; 95% confidence interval [CI], 1.37-1.91), history of MDD prior to injury (but not at the time of injury) (RR, 1.54; 95% CI, 1.31-1.82), age (RR, 0.61; 95% CI, 0.44-0.83 for > or = 60 years vs 18-29 years), and lifetime alcohol dependence (RR, 1.34; 95% CI, 1.14-1.57). Those with MDD were more likely to report comorbid anxiety disorders after TBI than those without MDD (60% vs 7%; RR, 8.77; 95% CI, 5.56-13.83). Only 44% of those with MDD received antidepressants or counseling. After adjusting for predictors of MDD, persons with MDD reported lower quality of life at 1 year compared with the nondepressed group. Among a cohort of patients hospitalized for TBI, 53.1% met criteria for MDD during the first year after TBI. Major depressive disorder was associated with history of MDD and was an independent predictor of poorer health-related quality of life.


lncRNA TINCR attenuates the proliferation and invasion, and enhances the apoptosis of cutaneous malignant melanoma cells by regulating the miR‑424‑5p/LATS1 axis.
Cutaneous malignant melanoma (CMM) is responsible for ≥1/2 of skin cancer‑related mortalities. The aberrant expression of long non‑coding RNAs (lncRNAs) has been associated with the development of CMM. However, to the best of our knowledge, the role of the lncRNA TINCR ubiquitin domain containing (TINCR) in CMM has not been previously investigated, and thus, the current study aimed to evaluate this in vitro and in vivo. Reverse transcription‑quantitative PCR (RT‑qPCR) was used to analyze microRNA (miR)‑424‑5p expression, and RT‑qPCR and western blotting were used to measure TINCR, large tumor suppressor kinase 1 (LATS1), cellular communication network factor 2 (CTGF), cellular communication network factor 1 (CCN1) and AXL receptor tyrosine kinase (AXL) mRNA and protein expression levels, respectively. Cell Counting Kit‑8, flow cytometry and Transwell assays were used to detect the proliferation, apoptosis and invasion of CMM cell lines, respectively. The binding sites between TINCR and miR‑424‑5p were predicted using the miRDB database. A dual luciferase reporter assay and RT‑qPCR were used to identify the relationship between TINCR and miR‑424‑5p in CMM cell lines. The bioinformatics analysis revealed that TINCR was one of the most significantly downregulated lncRNAs in CMM, and advanced stage CMM tissues showed the greatest decrease in TINCR expression. Moreover, in the collected CMM tissues and tested cell lines of the current study, TINCR expression was found to be downregulated compared with the respective controls. Notably, TINCR overexpression inhibited the expression levels of CTGF, CCN1 and AXL, decreased the proliferation and invasion, and induced the apoptosis of CMM cell lines. In addition, a mutual binding association was identified between miR‑424‑5p and TINCR in CMM cells. LATS1, a target of miR‑424‑5p, was found to be positively regulated by TINCR. TINCR activated Hippo signaling and repressed the activity of Yes 1 associated transcriptional regulator by regulating LATS1 expression, while LATS1 knockdown reversed the effect of TINCR overexpression on CMM cells. Collectively, the findings of the present study suggested that TINCR may attenuate the progression of CMM by regulating the miR‑424‑5p/LATS1 signaling axis. These results indicated that TINCR may play a tumor suppressive role in CMM.


Effects of docosahexaenoic acid and eicosapentaoic acid supplementation on white matter integrity after repetitive sub-concussive head impacts during American football: Exploratory neuroimaging findings from a pilot RCT.
Repetitive sub-concussive head impacts (RSHIs) are common in American football and result in changes to the microstructural integrity of white matter. Both docosahexaenoic acid (DHA) and eicosapentaoic acid (EPA) supplementation exerted neuroprotective effects against RSHIs in animal models and in a prior study in football players supplemented with DHA alone. Here, we present exploratory neuroimaging outcomes from a randomized controlled trial of DHA + EPA supplementation in American football players. We hypothesized that supplementation would result in less white matter integrity loss on diffusion weighted imaging over the season. We conducted a double-blind placebo-controlled trial in 38 American football players between June 2019 and January 2020. Participants were randomized to the treatment (2.442 g/day DHA and 1.020 g/day EPA) or placebo group for five times-per-week supplementation for 7 months. Of these, 27 participants were included in the neuroimaging data analysis (n = 16 placebo; n = 11 DHA + EPA). Changes in white matter integrity were quantified using both voxelwise diffusion kurtosis scalars and deterministic tractography at baseline and end of season. Additional neuroimaging outcomes included changes in regional gray matter volume as well as intra-regional, edge-wise, and network level functional connectivity. Serum neurofilament light (NfL) provided a peripheral biomarker of axonal damage. No voxel-wise between-group differences were identified on diffusion tensor metrics. Deterministic tractography using quantitative anisotropy (QA) revealed increased structural connectivity in ascending corticostriatal fibers and decreased connectivity in long association and commissural fibers in the DHA+EPA group compared to the placebo group. Serum NfL increases were correlated with increased mean (ρ = 0.47), axial (ρ = 0.44), and radial (ρ = 0.51) diffusivity and decreased QA (ρ = -0.52) in the corpus callosum and bilateral corona radiata irrespective of treatment group. DHA + EPA supplementation did preserve default mode/frontoparietal control network connectivity (g = 0.96, p = 0.024). These exploratory findings did not provide strong evidence that DHA + EPA prevented or protected against axonal damage as quantified via neuroimaging. Neuroprotective effects on functional connectivity were observed despite white matter damage. Further studies with larger samples are needed to fully establish the relationship between omega-3 supplementation, RSHIs, and neuroimaging biomarkers. ClinicalTrials.gov-NCT04796207.


[Resource investigation about Tibetan medicine Rhodiola kirilowii].
The investigation aims to better understand the resource status of Rhodiola kirilowii, analysis the suitable habitat of wild Rh. kirilowii and protect the wild resources of Rh. Kirilowii, promoting the sustainable utilization of Rh. kirilowii resources. In this paper, we investigated the wild resources of Rh. kirilowii in 16 counties of Sichuan, Qinghai, Gansu and Yunnan by means of investigation and sampling investigation combined with interview. The results showed that the population densities of wild Rh. kirilowii in 4 provinces were very different and the reserve of wild resources decreased gradually in many areas. According to the survey results, the current total reserve of Rh. kirilowii in four provinces was about 1 100 t. The reserve of wild Rh. kirilowii in Sichuan province was the largest. Simultaneously, the Rh. kirilowii had a certain ecological value. We found that a sand control base with planting Rh. kirilowii was set up in Hongyuan County of Sichuan Aba Tibetan and Qiang Autonomous Prefecture. The investigation provides a scientific basis for the development and sustainable utilization of Rh. kirilowii resources.


In the line-up: deleted genes associated with DiGeorge/22q11.2 deletion syndrome: are they all suspects?
22q11.2 deletion syndrome (22q11DS), a copy number variation (CNV) disorder, occurs in approximately 1:4000 live births due to a heterozygous microdeletion at position 11.2 (proximal) on the q arm of human chromosome 22 (hChr22) (McDonald-McGinn and Sullivan, Medicine 90:1-18, 2011). This disorder was known as DiGeorge syndrome, Velo-cardio-facial syndrome (VCFS) or conotruncal anomaly face syndrome (CTAF) based upon diagnostic cardiovascular, pharyngeal, and craniofacial anomalies (McDonald-McGinn and Sullivan, Medicine 90:1-18, 2011; Burn et al., J Med Genet 30:822-4, 1993) before this phenotypic spectrum was associated with 22q11.2 CNVs. Subsequently, 22q11.2 deletion emerged as a major genomic lesion associated with vulnerability for several clinically defined behavioral deficits common to a number of neurodevelopmental disorders (Fernandez et al., Principles of Developmental Genetics, 2015; Robin and Shprintzen, J Pediatr 147:90-6, 2005; Schneider et al., Am J Psychiatry 171:627-39, 2014). The mechanistic relationships between heterozygously deleted 22q11.2 genes and 22q11DS phenotypes are still unknown. We assembled a comprehensive "line-up" of the 36 protein coding loci in the 1.5 Mb minimal critical deleted region on hChr22q11.2, plus 20 protein coding loci in the distal 1.5 Mb that defines the 3 Mb typical 22q11DS deletion. We categorized candidates based upon apparent primary cell biological functions. We analyzed 41 of these genes that encode known proteins to determine whether haploinsufficiency of any single 22q11.2 gene-a one gene to one phenotype correspondence due to heterozygous deletion restricted to that locus-versus complex multigenic interactions can account for single or multiple 22q11DS phenotypes. Our 22q11.2 functional genomic assessment does not support current theories of single gene haploinsufficiency for one or all 22q11DS phenotypes. Shared molecular functions, convergence on fundamental cell biological processes, and related consequences of individual 22q11.2 genes point to a matrix of multigenic interactions due to diminished 22q11.2 gene dosage. These interactions target fundamental cellular mechanisms essential for development, maturation, or homeostasis at subsets of 22q11DS phenotypic sites.


LM-GVP: an extensible sequence and structure informed deep learning framework for protein property prediction.
Proteins perform many essential functions in biological systems and can be successfully developed as bio-therapeutics. It is invaluable to be able to predict their properties based on a proposed sequence and structure. In this study, we developed a novel generalizable deep learning framework, LM-GVP, composed of a protein Language Model (LM) and Graph Neural Network (GNN) to leverage information from both 1D amino acid sequences and 3D structures of proteins. Our approach outperformed the state-of-the-art protein LMs on a variety of property prediction tasks including fluorescence, protease stability, and protein functions from Gene Ontology (GO). We also illustrated insights into how a GNN prediction head can inform the fine-tuning of protein LMs to better leverage structural information. We envision that our deep learning framework will be generalizable to many protein property prediction problems to greatly accelerate protein engineering and drug development.


Stepwise Two-Photon-Gated Photochemical Reaction in Photochromic [2.2]Paracyclophane-Bridged Bis(imidazole dimer).
Stepwise two-photon processes not only have great potential for efficient light harvesting but also can provide valuable insights into novel photochemical sciences. Here we have designed a [2.2]paracyclophane-bridged bis(imidazole dimer), a molecule that is composed of two photochromic units and absorbs two photons in a stepwise manner. The absorption of the first photon leads to the formation of a short-lived biradical species (half-life = 88 ms at 298 K), while the absorption of the additional photon by the biradical species triggers a subsequent photochromic reaction to afford a long-lived quinoid species. The short-lived biradical species and the long-lived quinoid species display significantly different absorption spectra and rates of the thermal back-reaction. The stepwise two-photon excitation process in this photochromic system can be initiated even by incoherent continuous-wave light irradiation, indicating that this two-photon reaction is highly efficient. Our molecule based on the bridged bis(imidazole dimer) unit should be a good candidate for multiphoton-gated optical materials.


Clinical perspectives: practical insights from clinical experience with cabazitaxel in Australia.
Prostate cancer, and in particular the management options for patients with metastatic castration-resistant prostate cancer (mCRPC), remains an important health issue. The approval of cabazitaxel provides a new treatment option for patients who have progressed despite docetaxel therapy. Clinical use of cabazitaxel in mCRPC is based on the results of the TROPIC study, which demonstrated an approximately 30% reduction in the risk of death compared with mitoxantrone. In this paper, we draw on accumulating clinical experience with cabazitaxel in Australia to discuss important questions such as how to select the right patient for treatment and how to proactively prevent and manage toxicities associated with this therapy. Recognizing the need for a multidisciplinary approach to patient care, opinion and insight has been sought from medical oncologists, nurses and pharmacists. While lack of trial data means that a number of questions remain unanswered, local clinical experience has helped to guide treatment decisions and refine management protocols. Appropriate patient selection, careful ongoing monitoring and proactive management of adverse events will ensure optimal treatment of patients. Nurses in particular play an important role in educating patients and identifying patients at increased risk of developing adverse events with cabazitaxel. The oncology team must work together to educate patients; taking a proactive approach to issues such as adverse events will help maximize the clinical outcome of cabazitaxel treatment and impact positively on the patient's quality of life.


Atrial ejection force in systemic autoimmune diseases.
Systemic autoimmune disorders may affect several organs, including the heart. We analyzed two-dimensional and pulsed Doppler echocardiograms of patients (n = 37) with systemic lupus erythematosus (SLE, n = 24) or rheumatoid arthritis (RA, n = 13) to determine whether atrial ejection force (AEF) could represent a suitable parameter for detecting left ventricular filling abnormalities in SLE and RA. In both patient subgroups, AEF was significantly higher than in healthy controls (n = 40) matched for gender and age (14.0 +/- 5.4 vs. 11.0 +/- 3.5 kdyn, p < 0.01). Because conventional echocardiographic parameters of left ventricular function failed to detect such a difference, AEF might serve as an additional sensitive parameter for detecting left ventricular diastolic filling abnormalities early in the course of a systemic autoimmune disease.


Performance of the linear ion trap Orbitrap mass analyzer for qualitative and quantitative analysis of drugs of abuse and relevant metabolites in sewage water.
This work illustrates the potential of liquid chromatography coupled to a hybrid linear ion trap Fourier Transform Orbitrap mass spectrometer for the simultaneous identification and quantification of 24 drugs of abuse and relevant metabolites in sewage water. The developed methodology consisted of automatic solid-phase extraction using Oasis HLB cartridges, chromatographic separation of the targeted drugs, full-scan accurate mass data acquisition under positive electrospray ionization mode over an m/z range of 50-600Da at a resolution of 30,000 FWHM and simultaneous MS(n) measurements to obtain information of fragment ions generated in the linear ion trap. Accurate mass of the protonated molecule, together with at least one nominal mass product ion and retention time allowed the confident identification of the compounds detected in these complex matrices. In addition to the highly reliable qualitative analysis, Orbitrap analyzer also proved to have satisfactory potential for quantification at sub-ppb analyte levels, a possibility that has been very little explored in the literature until now. The limits of quantification ranged from 4 to 68ngL(-1) in influent sewage water, and from 2 to 35ngL(-1) in effluent, with the exception of MDA, morphine and THC that presented higher values as a consequence of the high ionization suppression in this type of samples. Satisfactory recoveries (70-120%) and precision (<30%) for the overall procedure were obtained for all compounds with the exception of meta-chlorophenylpiperazine, methylphenidate and ketamine. Isotope-labelled internal standards were added to sewage samples as surrogates in order to correct for matrix effects and also for possible losses during sample treatment. The methodology developed was applied to sewage water samples from the Netherlands (influent and effluent), and the results were compared with those obtained by LC-MS/MS with triple quadrupole. Several drugs of abuse could be identified and quantified, mainly MDMA, benzoylecgonine, codeine, oxazepam and temazepam. Orbitrap also showed potential for retrospective investigation of ketamine metabolites in the samples without the need of additional analysis.


Evaluation of an automated hemostasis testing analyzer, the Trombolyzer Combi.
We evaluated the Trombolyzer Combi (Behnk Elektronik, Norderstedt, Germany), an automated hemostasis analyzer, in a clinical setting. Determination of prothrombin time (PT), activated partial prothrombin time (APTT), fibrinogen (FIB) and antithrombin (AT) were performed using Organon Teknika reagents. Determination of PT, APTT and FIB on a KC4 (Amelung, Germany) using Dade reagent (Dade Behring, The Netherlands) and determination of AT on a Hitachi 912 using Chromogenix reagent (Nodia, The Netherlands) were used as reference methods. Within-run and total precision of the tests were determined by measuring pooled plasma samples at various levels in duplicate twice daily for twenty days. For all tests the within-run and total precision of the Trombolyzer Combi was comparable or superior to the reference methods. Methods comparison was performed with 100 patient samples for PT, APTT and FIB and with 50 patient samples for AT. The correlation coefficients between the Trombolyzer Combi values and the results from the reference methods were between 0.87 and 0.98. No effect of hemolysis on the determination of the studied parameters was detected. However, bilirubinemia above 260 micromol/L and triglycerides above 9 mmol/L resulted in erroneous test results. In conclusion, it is shown that the Trombolyzer Combi performs equivalently or better than the reference methods and can be used as a state-of-the-art hemostasis analyzer in a clinical laboratory.


Development of a Virtual Human for Supporting Tobacco Cessation During the COVID-19 Pandemic.
People who consume tobacco are at greater risk of developing severe COVID-19. Unfortunately, the COVID-19 pandemic reduced the accessibility of tobacco cessation services as a result of necessary social restrictions. Innovations were urgently needed to support tobacco cessation during the pandemic. Virtual humans are artificially intelligent computer agents with a realistic, humanlike appearance. Virtual humans could be a scalable and engaging way to deliver tobacco cessation information and support. Florence, a virtual human health worker, was developed in collaboration with the World Health Organization to remotely support people toward tobacco cessation during the COVID-19 pandemic. Florence delivers evidence-based information, assists with making quit plans, and directs people to World Health Organization-recommended cessation services in their country. In this viewpoint, we describe the process of developing Florence. The development was influenced by a formative evaluation of data from 115 early users of Florence from 49 countries. In general, Florence was positively perceived; however, changes were requested to aspects of her design and content. In addition, areas for new content were identified (eg, for nonsmoker support persons). Virtual health workers could expand the reach of evidence-based tobacco cessation information and personalized support. However, as they are a new innovation in tobacco cessation, their efficacy, feasibility, and acceptability in this application needs to be evaluated, including in diverse populations.


The Helicobacter pylori Genome Project: insights into H. pylori population structure from analysis of a worldwide collection of complete genomes.
Helicobacter pylori, a dominant member of the gastric microbiota, shares co-evolutionary history with humans. This has led to the development of genetically distinct H. pylori subpopulations associated with the geographic origin of the host and with differential gastric disease risk. Here, we provide insights into H. pylori population structure as a part of the Helicobacter pylori Genome Project (HpGP), a multi-disciplinary initiative aimed at elucidating H. pylori pathogenesis and identifying new therapeutic targets. We collected 1011 well-characterized clinical strains from 50 countries and generated high-quality genome sequences. We analysed core genome diversity and population structure of the HpGP dataset and 255 worldwide reference genomes to outline the ancestral contribution to Eurasian, African, and American populations. We found evidence of substantial contribution of population hpNorthAsia and subpopulation hspUral in Northern European H. pylori. The genomes of H. pylori isolated from northern and southern Indigenous Americans differed in that bacteria isolated in northern Indigenous communities were more similar to North Asian H. pylori while the southern had higher relatedness to hpEastAsia. Notably, we also found a highly clonal yet geographically dispersed North American subpopulation, which is negative for the cag pathogenicity island, and present in 7% of sequenced US genomes. We expect the HpGP dataset and the corresponding strains to become a major asset for H. pylori genomics.


[Cartography of healthcare for pregnant women].
This work uses cartography as a method for mapping the trajectory of primary healthcare provided to pregnant women. The scope of the study comprises 9 Basic Healthcare Units located in the city of Juazeiro do Norte in the State of Ceará. In all, fifteen women in the 37th to 39th week of pregnancy were selected. Interviews were conducted with these women during the period from January to June 2010. The cartographic findings were depicted in stages in the flowchart, which exposed lacunas in prenatal healthcare, such as the low number of oncotic cytology exams conducted and the lack of educational counseling. Nevertheless, in the interviews, a significant number of pregnant women expressed satisfaction with the prenatal care provided. The good relationships developed between the healthcare professionals and the pregnant women were the main reason that led them to continue the treatment. This fact reinforces the importance of dialogue between these two actors for the success of prenatal healthcare.


Experimental model for transforaminal endoscopic spine.
To validate the porcine spine as a model for learning and practicing transforaminal percutaneous endoscopic lumbar procedures (TF-PELP). TF-PELP was performed in three porcine cadaver lumbar spine levels. Anatomical features of the current cadaver were compared to human and porcine spines. Performance and documentation of endoscopic procedures were described. This study shows that this representative animal model reflects anatomical characteristics of the human spine. Transforaminal approaches were successfully completed. Although lower disc heights make disc puncture more difficult, the outside-in technique is feasible and more useful to identify anatomical parameters and to practice different surgical steps and maneuvers. This is an effective and representative model for learning and practicing this procedure. Difficulties of the procedure, as well as the differences compared to the human spine, were described.


Interactive streaming of stored multiview video using redundant frame structures.
While much of multiview video coding focuses on the rate-distortion performance of compressing all frames of all views for storage or non-interactive video delivery over networks, we address the problem of designing a frame structure to enable interactive multiview streaming, where clients can interactively switch views during video playback. Thus, as a client is playing back successive frames (in time) for a given view, it can send a request to the server to switch to a different view while continuing uninterrupted temporal playback. Noting that standard tools for random access (i.e., I-frame insertion) can be bandwidth-inefficient for this application, we propose a redundant representation of I-, P-, and "merge" frames, where each original picture can be encoded into multiple versions, appropriately trading off expected transmission rate with storage, to facilitate view switching. We first present ad hoc frame structures with good performance when the view-switching probabilities are either very large or very small. We then present optimization algorithms that generate more general frame structures with better overall performance for the general case. We show in our experiments that we can generate redundant frame structures offering a range of tradeoff points between transmission and storage, e.g., outperforming simple I-frame insertion structures by up to 45% in terms of bandwidth efficiency at twice the storage cost.